Antithrombotic azacycloalkylalkanoyl peptides and pseudopeptides

ABSTRACT

The present invention relates to azacycloalkylalkanoyl peptides and pseudopeptides which inhibit platelet aggregation and thrombus formation thereby being useful in the prevention and treatment of thrombosis associated with disease states such as myocardial infarction, stroke, peripheral arterial disease, and disseminated intravascular coagulation, to methods for the prevention or treatment of thrombosis in a mammal in need of such therapy comprising the administration of a therapeutically effective amount of such compounds, and to pharmaceutical compositions comprising such compounds.

This application is a 371 of PCT/US94/12135, filed Oct. 17, 1994, whichis a continuation-in-part application of co-pending U.S. applicationSer. No. 08/138,820, filed Oct. 15, 1993 abandoned.

BACKGROUND OF INVENTION

1. Field of Invention

This invention relates to compounds having antithrombotic activity. Moreparticularly, the invention relates to azacycloalkylalkanoyl peptidesand pseudopeptides that inhibit platelet aggregation and thrombusformation in mammals and which are useful in the prevention andtreatment of thrombosis associated with disease states such asmyocardial infarction, stroke, peripheral arterial disease anddisseminated intravascular coagulation.

Haemostasis, the biochemistry of blood coagulation, is an extremelycomplex phenomena whereby normal whole blood and body tissuespontaneously arrest bleeding from injured blood vessels. Effectivehaemostasis requires the combined activity of vascular, platelet andplasma factors as well as a controlling mechanism to prevent excessiveclotting. Defects, deficiencies, or excesses of any of these componentscan lead to hemorrhagic or thrombotic consequences.

Platelet adhesion, spreading and aggregation on extracellular matricesare central events in thrombus formation. These events are mediated by afamily of adhesive glycoproteins, i.e., fibrinogen, fibronectin, and vonWillebrand factor. Fibrinogen is a co-factor for platelet aggregation,while fibronectin supports platelet attachments and spreading reactions,and von Willebrand factor is important in platelet attachment to andspreading on subendothelial matrices. The binding sites for fibrinogen,fibronectin and von Willebrand factor have been located on the plateletmembrane protein complex known as glycoprotein IIb/IIIa.

Adhesive glycoproteins, like fibrinogen, do not bind with normal restingplatelets. However, when a platelet is activated with an agonist such asthrombin or adenosine diphosphate, the platelet changes its shape,perhaps making the GPIIb/IIIa binding site accessible to fibrinogen.Compounds within the scope of the present invention block the fibrinogenreceptor, thus inhibiting platelet aggregation and subsequent thrombusformation and when administered in the form of pharmaceuticalcompositions comprising such compounds are useful for the prevention andtreatment of thrombogenic diseases, such as myocardial infarction,stroke, peripheral arterial disease and disseminated intravascularcoagulation.

2. Reported Developments

It has been observed that the presence of Arg-Gly-Asp (RGD) is necessaryin fibrinogen, fibronectin and von Willebrand factor for theirinteraction with the cell surface receptor (Ruoslahti E., Pierschbacher,Cell 1986, 44, 517-18). Two other amino acid sequences also seem to takepart in the platelet attachment function of fibrinogen, namely, theGly-Pro-Arg sequence, and the dodecapeptide,His-His-Leu-Gly-Gly-Ala-Lys-Gln-Ala-Gly-Asp-Val sequence. Smallsynthetic peptides containing the RGD or dodecapeptide have been shownto bind to the platelet GPIIb/IIIa receptor and competitively inhibitbinding of fibrinogen, fibronectin and von Willebrand factor as well asinhibit aggregation of activated platelets (Plow, et al., Proc. Natl.Acad. Sci. USA 1985, 82, 8057-61; Ruggeri, et al., Proc. Natl. Acad.Sci. USA 1986, 5708-12; Ginsberg, et al., J. Biol. Chem. 1985, 260,3931-36; and Gartner, et al., J. Biol. Chem. 1987, 260,11,891-94).

Indolyl compounds containing guanidinoalkanoyl- and guandinoalkenoyl-aspartyl moieties are reported to be platelet-aggregation inhibitors byTjoeng, et al., U.S. Pat. Nos. 5,037,808 and 4,879,313.

U.S. Pat. No. 4,992,463 (Tjoeng, et al.), issued Feb. 12, 1991,discloses generically that a series of aryl and aralkyl guanidinoalkylpeptide mimetic compounds exhibit platelet aggregation inhibitingactivity and discloses specifically a series of mono- and dimethoxyphenyl peptide mimetic compounds and a biphenylalkyl peptide mimeticcompound.

U.S. Pat. No. 4,857,508 (Adams, et al.), issued Aug. 15,1989, disclosesgenerically that a series of guandinoalkyl peptide derivativescontaining terminal aralkyl substituents exhibit platelet aggregationinhibiting activity and discloses specifically a series of O-methyltyrosine, biphenyl, and naphthyl derivatives containing a terminal amidefunctionality.

Haverstick, D. M., et al., in Blood 66 (4), 946-952 (1985), disclosethat a number of synthetic peptides, including arg-gly-asp-ser andgly-arg-gly-asp-ser, are capable of inhibiting thrombin-induced plateletaggregation.

Plow, E. F., et al., in Proc. Natl. Acad. Sci. USA 79, 3711-3715 (1982),disclose that the tetrapeptide glycyl-L-prolyl-L-arginyl-L-prolineinhibits fibrinogen binding to human platelets.

French Application No. 86/17507, filed Dec. 15, 1986, discloses thattetra-, penta- and hexapeptide derivatives containing the-arg-gly-asp-sequence are useful as antithrombotics.

U.S. Pat. No. 4,683,291 (Zimmerman, et al.), issued Jul. 28, 1987,discloses that a series of peptides, comprised of from six to fortyamino acids, which contain the sequence -arg-gly-asp- are plateletbinding inhibitors.

European Application Publication No. 0 319 506, published Jun. 7, 1989,discloses that a series of tetra-, penta-, and hexapeptide derivativescontaining the -arg-gly-asp- sequence are platelet aggregationinhibitors.

Cyclic peptide analogues containing the moiety Gly-Asp are reported tobe fibrinogen receptor antagonists in U.S. Pat. No. 5,023,233.

Peptides and pseudopeptides containing amino-, guanidino-, imidizaloyl,and/or amidinoalkanoyl, and alkenoyl moieties are reported to beantithrombotic agents in pending U.S. applications Ser. Nos. 07/677,006,07/534,385, and 07/460,777 filed on Mar. 28, 1991, Jun. 7, 1990, andJan. 4, 1990, respectively, as well as in U.S. Pat. No. 4,952,562, andin International Application No. PCT/US90/05448, filed Sep. 25, 1990,all assigned to the same assignee as the present invention.

Peptides and pseudopeptides containing amino- and guanidino- alkyl- andalkenyl- benzoyl, phenylalkanoyl, and phenylalkenoyl moieties arereported to be antithrombotic agents in pending U.S. application Ser.No. 07/475,043, filed Feb. 5, 1990, and in International Application No.PCT/US91/02471, filed Apr. 11, 1991, published as InternationalPublication No. WO 92/13117 Oct. 29, 1992, assigned to the same assigneeas the present invention.

Alkanoyl and substituted alkanoyl azacycloalkylformyl aspartic acidderivatives are reported to be platelet aggregation inhibitors in U.S.Pat. No. 5,053,392, filed Dec. 1, 1989, and assigned to the sameassignee and having the same inventorship as the present invention.

N-subsituted azacycloalkylcarbonyl cyclic aminoacylaspartic acidderivatives are reported to be antithrombotics in U.S. Pat. No.5,064,814, filed Apr. 5, 1990 by the same inventors and assigned to thesame assignee as the present invention. Azacycloalkylformylglycylaspartic acid derivatives are reported to be antithrombotics in U.S.Pat. No. 5,051,405, filed Oct. 10, 1989, and assigned to the sameassignee as the present invention.

European Patent Application 0479,481, published Apr. 8, 1992, disclosesazacycloalkyalkanoyl glycyl aspartyl amino acids as fibrinogen receptorantagonists.

European Patent Application 0478,362, published Apr. 1, 1992, disclosesazacycloalkyalkanoyl peptidyl β-alanines as fibrinogen receptorantagonists.

The present invention relates to azacycloalkylalkanoyl peptides andpseudopeptides which inhibit platelet aggregation and thrombusformation.

SUMMARY OF THE INVENTION

Compounds of the present invention are described by Formula I ##STR1##wherein: A is -H, amidino, or substituted amidino;

B is alkyl, cycloalkyl, cycloalkylalkyl, alkylcycloalkyl,alkyycycloalkylalkyl, aryl, aralkyl, alkylaryl, or alkylaralkyl;##STR2## E is -H or, in combination with F, forms a 4-, 5-, 6-, or7-membered azacycloalkane ring,

F is the α-carbon side chain of a naturally occuring α-amino acid, -H,alkyl, cycloalkyl, cycloalkylalkyl, alkylcycloalkyl,alkylcycloalkylalkyl, aryl, substituted aryl, aralkyl, substituedaralkyl, heterocyclyl, substituted heterocyclyl, heterocyclylalkyl,substituted heterocyclylalkyl, or, in combination with E, forms a 4-,5-, 6-, or 7-membered azacycloalkane ring,

G is alkyl, cycloalkyl, cycloalkylalkyl, alkylcycloalkyl,alkylcycloalkylalkyl, aryl, substituted aryl, aralkyl, substitutedaralkyl, heterocyclyl, substituted heterocyclyl, heterocyclylalkyl,substituted heterocyclylalkyl, OR¹, or NR¹ R², where R¹ and R² areindependently -H, alkyl, cycloalkyl, cycloalkylalkyl, alkylcycloalkyl,alkylcycloalkylalkyl, aryl, aralkyl, alkylaryl, or alkylaralkyl, and

r is 0 or 1;

R is H-, alkyl, aryl, or aralkyl;

m is 1 to 5;

n is 0 to 6; and

p is 1 to 4;

or a pharmaceutically acceptable salt thereof.

Additionally, the present invention relates to pharmaceuticalcompositions comprising such compounds, and to methods of prevention ortreatment of thrombosis in a mammal in need of such therapy comprisingthe administration of such compounds and pharmaceutical compositions.

The present invention is characterized by the marked and prolongedantithrombotic activity of the compounds of Formula I, above, observedafter oral administration thereof.

DETAILED DESCRIPTION OF INVENTION

As used above, and throughout the description of this invention, thefollowing terms, unless otherwise indicated, shall be understood to havethe following meanings: ##STR3##

"Substituted amidino" means an amidino group N-substituted on one orboth nitrogens by one or more alkyl, cycloalkyl, cycloalkylalkyl,alkycycloalkyl, alkylcycloalkylalkyl, aryl or aralkyl groups.

"Alkyl" means a saturated aliphatic hydrocarbon group which may bestraight or branched and having about 1 to about 20 carbon atoms in thechain. Branched means that a lower alkyl group such as methyl, ethyl orpropyl is attached to a linear alkyl chain. Preferred straight orbranched alkyl groups are the "lower alkyl" groups which are those alkylgroups having from 1 to about 10 carbon atoms. Most preferred loweralkyl groups have from 1 to about 6 carbon atoms.

"Cycloalkyl" means a saturated carbocyclic group having one or morerings and having about 3 to about 10 carbon atoms. Preferred cycloalkylgroups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,cycloheptyl, and decahydronaphthyl.

"Cycloalkylalkyl means an alkyl group substituted with a cycloalkylgroup. Preferred cycloalkylalkyl groups include cyclopentylmethyl,cyclohexylmethyl, cyclohexylethyl,decahydronaphth-1-ylmethyl anddecahydronaphth-2-ylmethyl.

"Alkylcycloalkyl" means an cycloalkyl group substituted with an alkylgroup. Exemplary alkylcycloalkyl groups include 1-, 2-, 3-, or 4-methylor ethyl cyclohexyl.

"Alkylcycloalkylalkyl" means an alkyl group substituted by analkylcycloalkyl group. Exemplary alkylcycloalkyl groups include 1-, 2-,3-, or 4-methyl or ethyl cyclohexylmethyl or 1-, 2-, 3-, or 4-methyl orethyl cyclohexylethyl.

"Azacycloalkane" means a saturated aliphatic ring containing a nitrogenatom. Preferred azacycloalkanes include pyrollidine and piperidine.

"Naturally occuring α-amino acid" means glycine, alanine, valine,leucine, isoleucine, serine, threonine, phenylalanine, tyrosine,tryptophan, cysteine, methionine, proline, hydroxyproline, asparticacid, asparagine, glutamine, glutamic acid, histidine, arginine,ornithine, and lysine.

"α-carbon side chain of a naturally occuring α-amino acid" means themoiety which substitutes the x-carbon of a naturally occuring α-aminoacid. Exemplary α-carbon side chains of naturally occuring α-amino acidsinclude isopropyl, methyl, and carboxymethyl for valine, alanine, andaspartic acid, respectively.

"Aryl" means a phenyl or naphthyl group.

"Substituted aryl" means a phenyl or naphthyl group substituted by oneor more aryl group substitutents which may be the same or different,where "aryl group substituent" includes alkyl, alkenyl, alkynyl, aryl,aralkyl, hydroxy, alkoxy, aryloxy, aralkoxy, hydroxyalkyl, acyl, formyl,carboxy, alkenoyl, aroyl, halo, nitro, trihalomethyl, cyano,alkoxycarbonyl, aryloxycarbonyl, aralkoxycarbonyl, acylamino,aroylamino, carbamoyl, alkylcarbamoyl, dialkylcarbamoyl, arylcarbamoyl,aralkylcarbamoyl, alkylsulfonyl, alkylsulfinyl, arylsulfonyl,arylsulfinyl, aralkylsulfonyl, aralkylsulfinyl, or -NR_(a) R_(b) whereRa and R_(b) are independently hydrogen, alkyl, aryl, or aralkyl.

"Aralkyl" means an alkyl group substituted by an aryl radical. Preferredaralkyl groups include benzyl, naphth-1-ylmethyl naphth-2-ylmethyl, andphenethyl.

"Substituted aralkyl" means an aralkyl group substituted on the arylportion by one or more aryl group substituents.

"Heterocyclyl" means about a 4- to about a 15-membered monocyclic ormulticyclic ring system in which one or more of the atoms in the ring isan element other than carbon, for example nitrogen, oxygen, or sulfur.Preferred heterocyclyl groups include pyridyl, pyrimidyl, andpyrrolidyl.

"Substituted heterocyclyl" means a heterocyclyl group substitued by oneor more aryl group substituents.

"Heterocyclylalkyl" and "substituted heterocyclylalkyl" means an alkylgroup which is substituted by a heterocyclyl and substitutedheterocyclyl group, respectively.

A preferred class of compounds of the present invention is described byFormula I wherein F is -H, alkyl, hydroxymethyl, 1-hydroxyethyl,mercaptomethyl, 2-methylthioethyl, carboxymethyl, 2-carboxyethyl,4-aminobutyl, 3-guanidinopropyl, cycloalkyl, cycloalkylalkyl,alkylcycloalkyl, alkylcycloalkylalkyl, aryl, substituted aryl, aralkyl,substituted aralkyl, heterocyclyl, substituted heterocyclyl,heterocyclylalkyl, substituted heterocyclylalkyl, or, in combinationwith E, forms a 4-, 5-, 6-, or 7-membered azacycloalkane ring, providedthat heterocyclylalkyl is other than indol-3-ylmethyl.

A more preferred class of compounds of the present invention isdescribed by the preferred class of compounds wherein F is -H, alkyl,hydroxymethyl, 1-hydroxyethyl, mercaptomethyl, 2-methylthioethyl,carboxymethyl, 2-carboxyethyl, 4-aminobutyl, 3-guanidinopropyl,cycloalkyl, cycloalkylalkyl, alkylcycloalkyl, alkylcycloalkylalkyl,aryl, substituted aryl, aralkyl, substituted aralkyl, or, in combinationwith E, forms a 4-, 5-, 6-, or 7-membered azacycloalkane ring.

A still more preferred class of compounds of the present invention isdescribed by the more preferred class of compounds wherein F is -H,alkyl, hydroxymethyl, 1-hydroxyethyl, mercaptomethyl, 2-methylthioethyl,carboxymethyl, 2-carboxyethyl, 4-aminobutyl, 3-guanidinopropyl,cycloalkyl, cycloalkylalkyl, alkylcycloalkyl, alkylcycloalkylalkyl, or,in combination with E, forms a 4-, 5-, 6-, or 7-membered azacycloalkanering.

A most preferred class of compounds of the present invention isdescribed by the still more preferred class of compounds wherein B isalkyl, cycloalkyl, cycloalkylalkyl, alkylcycloalkyl, oralkylcycloalkylalkyl

A special embodiment of the present invention is described by Formula II##STR4## wherein: A is -H or amidino,

B is alkyl, cycloalkyl, cycloalkylalkyl, alkylcycloalkyl,alkylcycloalkylalkyl, aryl, aralkyl, alkylaryl, or alkylaralkyl,

J is -H, alkyl, cycloalkyl, cycloalkylalkyl, alkylcycloalkyl,alkylcycloalkylalkyl, aryl, substituted aryl, aralkyl, substituedaralkyl,

L is OR¹, or NR¹ R², where R¹ and R² are independently -H, alkyl,cycloalkyl, cycloalkylalkyl, alkylcycloalkyl, alkylcycloalkylalkyl,aryl, aralkyl, alkylaryl, or alkylaralkyl,

m is 1 to 5,

n is 2 to 6, and

p is 1 or 2;

or a pharmaceutically acceptable salt thereof.

A more preferred special embodiment of the present invention isdescribed by the compounds of the special embodiment wherein

A is -H,

B is alkyl, cycloalkyl, cycloalkylalkyl, alkylcycloalkyl,alkylcycloalkylalkyl,

J is -H, alkyl, cycloalkyl, cycloalkylalkyl, alkylcycloalkyl, oralkylcycloalkylalkyl,

m is 3, and

n is 3 or 4.

A most preferred special embodiment of the special invention isdescribed by the compounds of the more preferred special embodimentwherein

A is -H,

B is alkyl,

J is alkyl, cycloalkyl, or cycloalkylalkyl,

R¹ and R² are independently -H, alkyl, cycloalkyl, cycloalkylalkyl,alkylcycloalkyl, alkylcycloalkylalkyl,

m is 3,

n is 3 or 4, and

p is 1.

Representative compounds of the present invention include:

N- N- N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl! valine,

N- N- N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-D-valine,

N- N- N-(3-(piperidin-4-yl)propanoyl)-N-ethylglycyl!aspartyl! valine,

N- N- N-(5-(piperidin-4-yl)pentanoyl)-N-ethylglycyl!aspartyl! valine,

N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-L-α-cyclohexylglycine,

N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-β-cyclohexylalanine,

N- N- N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl! norleucine,

N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-L-α-(2,2-dimethyl)prop-3-ylglycine,

N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-L-β-decahydronaphth-1-ylalanine,

N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-L-α-(2-cyclohexylethyl)glycine,

N- N- N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!phenylalanine,

N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-L-β-naphth-1-ylalanine,

N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-L-β-naphth-2-ylalanine,

N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-L-β-cyclohexylalanine amide,

N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-L-β-cyclohexylalanine, ethyl ester,

2-cyclohexyl-N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-ethylamine,

N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-L-β-cis-decahydronaphth-2-ylalanine,

3-Adamant-1-ylpropyl-N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartate,

N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-α-aminocyclohexanecarboxylicacid,

N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-β-cyclohexyl-D-alanine,

N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-β-decahydronaphth-1-ylalanine,

N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-β-cyclohexylalanineethyl amide,

N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-β-cyclooctylaianine,

N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-α-cyclohexylmethylethanolamine,

N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-β-cyclohexylmethylalanineamide,

N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-β-adamant-1-ylalanine,

N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-β-(1,2,3,4)-tetrahydronaphth-5-ylalanine,

N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-β-(4-cyclohexyl)cyclohexylalanine,

N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-β-cycloheptylalanine,

N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-β-cyclooctylalanineamide,

N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-α-cyclohexylpropylglycine,

N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-β-cyclooctylmethylalanine,

N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-β-cyclopentylalanine,and

N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-β-cyclohexylmethylalanineethyl ester, and

pharmaceutically acceptable salts thereof.

Compounds of the present invention contain asymmetric centers. Theseasymmetric centers may independently be in either the R or Sconfiguration. The present invention comprises the individualstereoisomers and mixtures thereof.

The compounds of the present invention may be useful in the form of thefree base or acid or in the form of a pharmaceutically acceptable saltthereof. All forms are within the scope of the invention.

Where the compound of the present invention is substituted with a basicmoiety, acid addition salts may be formed and are simply a moreconvenient form for use; and in practice, use of the salt forminherently amounts to use of the free base form. The acids which can beused to prepare the acid addition salts include preferably those whichproduce, when combined with the free base, pharmaceutically acceptablesalts, that is, salts whose anions are non-toxic to the animal organismin pharmaceutical doses of the salts, so that the beneficialantithrombotic properties inherent in the free base are not vitiated byside effects ascribable to the anions. Although pharmaceuticallyacceptable salts of said basic compounds are preferred, all acidaddition salts are useful as sources of the free base form even if theparticular salt, per se, is desired only as an intermediate product as,for example, when the salt is formed only for purposes of purification,and identification, or when it is used as intermediate in preparing apharmaceutically acceptable salt by ion exchange procedures.Pharmaceutically acceptable salts within the scope of the invention arethose derived from the following acids: mineral acids such ashydrochloric acid, sulfuric acid, phosphoric acid and sulfamic acid; andorganic acids such as acetic acid, citric acid, lactic acid, tartaricacid, malonic acid, methanesufonic acid, ethanesulfonic acid,benzenesulfonic acid, p-toluenesulfonic acid, cyclohexylsulfamic acid,quinic acid, and the like. The corresponding acid addition saltscomprise the following: hydrochloride, sulfate, phosphate, sulfamate,acetate, citrate, lactate, tartarate, malonate, methanesulfonate,ethanesulfonate, benzenesulfonate, p-toluenesulfonate,cyclohexylsulfamate and quinate, respectively.

The acid addition salts of the compounds of this invention are preparedeither by dissolving the free base in aqueous or aqueous-alcoholsolution or other suitable solvents containing the appropriate acid andisolating the salt by evaporating the solution, or by reacting the freebase and acid in an organic solvent, in which case the salt separatesdirectly or can be obtained by concentration of the solution.

Where the compound of the invention is substituted with an acidicmoiety, base addition salts may be formed and are simply a moreconvenient form for use; and in practice, use of the salt forminherently amounts to use of the free acid form. The bases which can beused to prepare the base addition salts include preferably those whichproduce, when combined with the free acid, pharmaceutically acceptablesalts, that is, salts whose cations are non-toxic to the animal organismin pharmaceutical doses of the salts, so that the beneficialantithrombotic properties inherent in the free acid are not vitiated byside effects ascribable to the cations. Pharmaceutically acceptablesalts within the scope of the invention are those derived from thefollowing bases: sodium hydroxide, potassium hydroxide, calciumhydroxide, aluminum hydroxide, lithium hydroxide, magnesium hydroxide,zinc hydroxide, ammonia, ethylenediamine, N-methyl-glucamine, lysine,arginine, ornithine, choline, N,N'-dibenzylethylenediamine,chloroprocaine, diethanolamine, procaine, N-benzylphenethylamine,diethylamine, piperazine, tris(hydroxymethyl)aminomethane,tetramethylammonium hydroxide, and the like.

Metal salts of compounds of the present invention may be obtained bycontacting a hydroxide, carbonate or similar reactive compound of thechosen metal in an aqueous solvent with the free acid form of thecompound. The aqueous solvent employed may be water or it may be amixture of water with an organic solvent, preferably an alcohol such asmethanol or ethanol, a ketone such as acetone, an aliphatic ether suchas tetrahydrofuran, or an ester such as ethyl acetate. Such reactionsare normally conducted at ambient temperature but they may, if desired,be conducted with heating.

Amine salts of compounds of the present invention may be obtained bycontacting an amine in an aqueous solvent with the free acid form of thecompound. Suitable aqueous solvents include water and mixtures of waterwith alcohols such as methanol or ethanol, ethers such astetrahydrofuran, nitrites such as acetonitrile, or ketones such asacetone. Amino acid salts may be similarly prepared.

Compounds of this invention may be prepared in accordance with thereaction sequences described below, or can be prepared by methods knownin the art. The starting materials used in the preparation of compoundsof this invention are known or are commercially available, or can beprepared by known methods or by specific reaction schemes describedherein.

The compounds of the present invention may be readily prepared bystandard solid phase or solution phase peptide synthesis proceduresusing starting materials and/or readily available intermediates fromchemical supply companies such as Aldrich or Sigma, (H. Paulsen, G.Merz, V. Weichart, "Solid-Phase Synthesis of O-Glycopeptide Sequences",Angew. Chem. Int. Ed. Engl. 27 (1988); H. Mergler, R. Tanner, J.Gosteli, and P. Grogg, "Peptide Synthesis by a Combination ofSolid-Phase and Solution Methods I: A New Very Acid-Labile Anchor Groupfor the Solid-Phase Synthesis of Fully Protected Fragments. Tetrahedronletters 29, 4005 (1988); Merrifield, R. B., "Solid Phase PeptideSynthesis after 25 Years: The Design and Synthesis of Antagonists ofGlucagon", Makromol. Chem. Macromol. Symp. 19, 31 (1988)).

A preferred method of preparing compounds of the present invention is bythe solution phase method depicted in Scheme I, below. ##STR5## whereinA, B, E, F, G, R, m, n, p, and r are as defined hereinabove; A', E', F',G', and R' are A, B, E, F, G, and R, respectively, or are protectedanalogues thereof, or precursor substituents thereto; and

P₁, P₂, and P₃ are amino acid protecting groups.

The compounds of the present invention are available generally byinitially coupling the appropriate amino acid or other appropriate Zgroup precursor, where Z is as defined hereinabove, which contains afree primary or secondary amine to the free carboxylic acid portion of aprotected derivative of aspartic acid.

The functional groups of aspartic acid or any functional groups of the Zgroup precursor which are not to be coupled are protected wherenecessary by blocking groups to prevent cross reaction during thecoupling procedure, as are the amino acid derivatives andazacycloalkylalkanoic acid derivatives used in subsequent syntheticsteps. These blocking groups include N-α-tertiary butyloxycarbonyl(BOC), benzyloxycarbonyl (CBZ), benzyl, methyl, t-butyl,9-fluorenylmethyloxycarbonyl (FMOC), 2-(trimethylsilyl)ethyl, and4-methoxy-2,3,6-trimethylbenzenesulfonyl.

A preferred protected derivative of aspartic acid is BOC aspartic acidβ-benzyl ester. Coupling is done by methods known in the art. Apreferred method for carrying out the coupling is combining the amineand carboxylic acid in an appropriate aprotic organic solvent, forexample methylene chloride or dimethylformamide (DMF), in the presenceof appropriate coupling agents. A preferred coupling agent is isopropylchloroformate in the presence of N-methylpiperidine. Another preferredcoupling agent is 1-(3-dimethylaminopropyl)-3-ethylcarbodiimidehydrochloride (EDC) in the presence of 1-hydroxybenzotriazole (HOBT) andtriethylamine. Still another preferred coupling agent isbis(2-oxo-3-oxazolidinyl)-phosphonic chloride (BOP-Cl) in the presenceof triethylamine.

The resulting protected product is selectively deprotected by knownmethods to give the N-terminal free amine of the aspartic acid moiety. Apreferred method for removing the BOC group is treatment withtrifluoroacetic in an aprotic organic solvent, for example, methylenechloride.

The resulting deprotected product is then coupled with the appropriatelyN-protected N-substituted glycine or β-alanine derivative having a freecarboxyl group. The resulting producted is then N-deprotected. Theresulting free amine is then coupled with the appropriate protectedazacycloalkylalkanoic acid and this product deprotected by known methodsto give the final product.

In another preferred method, the compounds of the present invention maybe prepared by solid phase methods well known in the art. In the solidmethod the C-terminal residue is bound at the carboxyl portion to aninsoluble resin, for example the residue may be bound as ap-alkoxybenzyl alcohol resin ester. In a manner which is similar to thesolution phase method the protected amino acid or other residues areadded one at a time until the total sequence has been built up on theresin. The compound is then deprotected and released from the resin bystandard methods to give the final compound.

During the preparation of compounds of the present invention, orintermediates thereto, it may also be desirable or necessary to preventcross-reaction between chemically active substituents other than thosepresent on naturally occuring or other amino acids. The substituents maybe protected by standard blocking groups which may subsequently beremoved or retained, as required, by known methods to afford the desiredproducts or intermediates (see, for example, Green, "Protective Groupsin Organic Synthesis", Wiley, N.Y., 1981). Selective protection ordeprotection may also be necessary or desirable to allow conversion orremoval of existing substituents, or to allow subsequent reaction toaffort the final desired product.

The invention is further explained by the following illustrativeexamples. In the following examples, unless otherwise indicated, α-aminoacids which have the possibility of having chiral α-carbons are in the Lconfiguration.

Unless otherwise indicated, reported mass spectral analysis data are LowResolution Fast Atom Bombardment performed on a VG 70SE with"calculated" values being (M+H)⁺. Nuclear magnetic resonance spectraldata is obtained on a Brucker ACF 300, in D₂ O. Flash chromatography isdone on silica gel. High performance liquid chromatography (HPLC) isdone on a Dynamax 60 Å, 8μ C-18 Reverse Phase column.

EXAMPLE 1

N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-β-cyclohexylalanine##STR6##A. β-cyclohexylalanine (1.12 g) is dissolved in of methanol (50ml) and hydrogen chloride gas is bubbled through the solution for about15 minutes. The solution is evaporated in vacuo and toluene azeotropedfrom the residue to give β-cyclohexyl-L-alanine methyl ester as thehydrochloride salt.

B. BOC-L-aspartic acid β-benzyl ester (1.27 g) is dissolved in methylenechloride (20 ml). The solution is cooled to 0° C. and N-methylpiperidine (0.48 ml) is added followed by isopropyl chloroformate(3.94ml). The solution is stirred at 0° C. for about two minutes andβ-cyclohexyl-L-alanine methyl ester hydrochloride (0.88 g) is added. Thesolution is allowed to warm to room temperature and stirred overnight.The solution is evaporated in vacuo and the residue partitionedbetweenethyl acetate (200 ml) and 1N hydrochloric acid (HCl) (50 ml). Theorganic layer is washed with 1N HCl, saturated sodium bicarbonatesolution, brine, dried over magnesium sulfate, filtered and evaporatedin vacuo to give N- BOC-L-aspartyl (β-benzylester)!-β-cyclohexyl-L-alanine, methyl ester.

C. N- BOC-L-aspartyl (β-benzyl ester)!-β-cyclohexylalanine, methyl ester(2.01 g) is dissolved in methylene chloride (15 ml). The solution iscooled to 0° C. and trifluoroacetic acid (5 ml) is added over a periodof about 1 minute. The solution is stirred at 0° C. for two hours,evaporated in vacuo and residue taken up into ethyl acetate and theorganic solution washed with saturated sodium bicarbonate solution untilthe washes are basic. The organic solution is washed with brine, driedover magnesium sulfate, filtered and evaporated in vacuo to giveL-aspartyl (β-benzyl ester)-β-cyclohexyl-L-alanine methyl ester.

D. Using essentially the coupling procedure of Example 1B, hereinabove,followed by essentially the deprotection procedure of Example 1C,hereinabove, N-ethylglycyl-L-aspartyl (β-benzylester)-L-β-cyclohexyl-L-alanine methyl ester is prepared from L-aspartyl(β-benzyl ester)-β-cyclohexyl-L-alanine methyl ester andN-BOC-N-ethylglycine.

E. 4-pyridine acetic acid (10 g) and platinum oxide (1.0 g) are combinedinacetic acid (100 ml) and the mixture shaken under hydrogen at 50 psifor about 18 hours. The mixture is filtered and the solution evaporatedin vacuo and toluene azeotroped from the residue to give2-(piperidin-4-yl)acetic acid.

F. 2-(piperidin-4-yl)acetic acid (11.6 g) is dissolved in 1N aqueoussodiumhydroxide solution (200 ml) and the solution cooled to 0° C. Asolution of di-tert-butyl dicarbonate (18.0 g) in tetrahydofuran (THF)(100 ml) is added dropwise and the mixture allowed to warm to roomtemperature and stirred for about 18 hours. The mixture is evaporated invacuo to remove THF and the residue is taken up into water and washedwithethyl acetate. Ethyl acetate is added to the aqueous layer and themixture acidified with 1N HCl. The organic layer is separated and theaqueous layer extracted with ethyl acetate. The combined organicportions are washed with water, brine, dried over magnesium sulfate,filtered, and evaporated in vacuo to give N-BOC-2-(piperidin-4-yl)aceticacid.

G. N-BOC-2-(piperidin-4-yl)acetic acid (15.8 g) is dissolved in THF (150ml) and 1M borane/THF (70 ml) is added dropwise. The solution is stirredat room temperature for about 20 hours and 1N sodium hydroxide solution(200 ml) is added dropwise. The THF is evaporated in vacuo and theaqueousresidue extracted with ethyl acetate. The ethyl acetate solutionis washed with water, dried over sodium sulfate, filtered, andevaporated in vacuo to give N-BOC-2-(piperidin-4-yl)ethanol.

H. A solution of oxalyl chloride (11.8 g) in methylene chloride (180 ml)iscooled to -78° C. and dimethyl sulfoxide (DMSO) (8.9 ml) is addeddropwise. The solution is stirred at -78° C. for about 3 minutes and asolution of N-BOC-2-(piperidin-4-yl)ethanol (14.3 g) in methylenechloride (250 ml) is added over a period of about 10 minutes. Thesolutionis stirred for about 1 hour and N-methyl morpholine (21.6 g) isadded over a period of about 15 minutes. The solution is allowed to warmto room temperature and, after about 30 minutes, methyl(triphenylphosphoranylidene) acetate (68.6 g) is added. The solution isstirred at room temperature for about 18 hours, evaporated in vacuo andthe residue taken up in ethyl acetate. The ethyl acetate solution iswashed with water, 5% HCl, 5% sodium hypochlorite solution, water,brine, dried over sodium sulfate, filtered and evaporated in vacuo togive methyl4-(N-BOC-piperidin-4-yl)trans-crotonate.

I. Methyl 4-(N-BOC-piperidin-4-yl)trans-crotonate (11.5 g) is dissolvedin methanol (200 ml) and 10% palladium/carbon (3 g) is added and themixture shaken under hydrogen at 50 psi for 18 hours. The mixture isfiltered, fresh catalyst added to the solution, and hydrogenationrepeated. The mixture is filtered and evaporated in vacuo to give methyl4-(N-BOC-piperidin-4-yl) butyrate.

J. To a mixture of 1N aqueous sodium hydroxide (100 ml) and methanol(200 ml) is added methyl 4-(N-BOC-piperidin-4-yl) butyrate (10.1 g) andthe mixture stirred at room temperature for about 18 hours. The mixtureis evaporated in vacuo, diluted with water, and washed with ether. Theaqueous portion is acidified with 5% HCl, extracted with ethyl acetateandthe organic solution washed with water, brine, dried over sodiumsulfate, filtered, and evaporated in vacuo to give4-(N-BOC-piperidin-4-yl) butyricacid.

K. A solution of 4-(N-BOC-piperidin-4-yl) butyric acid (0.91 g) inmethylene chloride (50 ml) is cooled to 0° C. andbis(2-oxo-3-oxazolidinyl)phosphinic chloride (BOP-Cl) (0.86 g) andtriethylamine (0.47 ml) are added. The solution is stirred at 0° C.forabout 10 minutes and N-ethylglycyl-L-aspartyl (β-benzylester)-L-β-cyclohexyl-L-alanine methyl ester (1.52 g) in a minimumofmethylene chloride is added followed by dropwise addition oftriethylamine (0.47 ml) in methylene chloride over a period of about 15minutes. The mixture is stirred at 0° C. for about 1 hour, and at roomtemperature for about 18 hours. The mixture is evaporated in vacuo andtheresidue taken up into ethyl acetate. The organic solution is washedwith 1NHCl, saturated sodium bicarbonate solution, brine, dried overmagnesium sulfate, filtered,evaporated in vacuo and the residue purifiedby flash chromatography, eluting with 60% ethyl acetate in hexanes, togive N- N-N-(4-(N-BOC-piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl(β-benzylester)!-β-cyclohexylalanine, methyl ester.

L. N- N-N-(4-(N-BOC-piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl(β-benzylester)!-β-cyclohexylalanine, methyl ester (1.79 g) is dissolved inmethanol (40 ml) and 10% palladium/carbon (0.25 g) is added. The mixtureis shaken under hydrogen at 50 psi for about 18 hours. The mixtureisfiltered through a Celite pad and the filtrate evaporated in vacuo togive N- N-N-(4-(N-BOC-piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-β-cyclohexylalanine,methyl ester. The ester is dissolved in methanol (20 ml)and 1N aqueoussodium hydroxide solution (10 ml) is added. The mixture is stirred atroom temperature for about 4 hours, diluted with water (25 ml),andacidified to pH 2 with 1N HCl. The mixture is extracted with ethylacetate (3×100 ml) and the ethyl acetate solution dried over magnesiumsulfate, filtered and evaporated in vacuo to give N- N-N-(4-(N-BOC-piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-β-cyclohexylalanine.

M. N- N-N-(4-(N-BOC-piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-β-cyclohexylalanine(1.39 g) is dissolved in methylene chloride (15 ml) and the solutioncooled to 0° C. Trifluoroacetic acid (5 ml) is added and the solutionstirred at 0° C. for about 2.5 hours. The solutionis evaporated in vacuoand the residue diluted with water, frozen, and lyophilized. The residueis purified by reverse phase HPLC, eluting with agradient of 40% to 80%methanol in water, containing 0.1% trifluoroacetic. The appropriatefractions are combined, and lyophilized to give N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-β-cyclohexylalanineas the trifluoroacetate salt. M.S., Calc'd: 525, Found: 525; NMR,δ=4.58-4.48 (m, 1H), 4.35-4.22 (m, 1H), 3.88 (s, 2H), 3.32 (q, 2H),3.28-3.10 (m, 2H), 2.88-2.60 (m, 4H), 2.33 (t, 2H), 1.85-1.70 (m, 2H),1.62-1.35 (m, 10H), 1.30-1.06 (m, 5H), 1.04-0.65 (m, 8H).

Using essentially the procedures of Example 1 above, the followingcompounds are prepared from the appropriate starting materials.

EXAMPLE 2

N- N- N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl! valine##STR7##

M.S., Calc'd: 471, Found: 471; NMR, δ=4.15-4.05 (m, 2H), 3.90 (s, 2H),3.30 (q, 2H), 3.30-3.15 (m, 2H), 2.90-2.60 (m, 4H), 2.33 (t, 2H), 2.05(q, 1H), 1.85-1.72 (m, 2H), 1.55-1.35 (m, 3H), 1.30-1.08 (m, 4H), 1.02(t, 3H), 0.70 (d,6H).

EXAMPLE 3

N- N- N-(4-piperidin-4-yl)-N-ethylglycyl!aspartyl!phenylalanine H##STR8##

M.S., Calc'd: 519, Found: 519; NMR, δ=7.20-6.95 (m, 5H), 4.55-4.35 (m,2H), 3.73 (s, 2H), 3.30-2.40 (m, 10H), 2.25 (t, 2H), 1.75-1.60 (m, 2H),1.4-1.25 (m, 3H), 1.20-0.97 (m, 4H), 0.92 (t, 3H).

EXAMPLE 4

N- N- N-4-(piperidin-4-yl)butanoyl)-N-ethylclycyl!aspartyl!-D-valine##STR9##

M.S., Calc'd: 471, Found: 471; NMR, δ=4.10-3.98 (m, 2H), 3.85 (s, 2H),3.28 (q, 2H), 3.23-3.10 (m, 2H), 2,82-2.58 (m, 4H), 2.28 (t, 2H), 2.02(q, 1H), 1.80-1.65 (m, 2H), 1.50-1.28 (m, 3H), 1.25-1.00 (m, 4H), 0.95(t, 3H), 0.78-0.65 (m, 6H).

EXAMPLE 5

N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-L-α-2,2-dimethyl)prop3-ylglycine ##STR10##

M.S., Calc'd: 499, Found: 499; NMR, δ=4.63-4.55 (m, 1H), 4.30-4.20 (m,1H), 3.88 (s, 2H), 3.33 (q, 2H), 3.30-3.15 (m, 2H), 2.88-2.60 (m,4H),2.35 (t, 2H), 1.85-1.75 (m, 2H), 1.73-1.35 (m, 5H), 1.30-1.08 (m,4H), 1.03(t, 3H), 0.78 (s, 9H).

EXAMPLE 6

N- N- N-(4-piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!norleucine##STR11##

M.S., Calc'd: 485, Found: 485; NMR, δ=4.58-4.50 (m, 1H), 4.20-4.10 (m,1H), 3.85 (s, 2H), 3.32 (q, 2H), 3.25-3.10 (m, 2H), 2.85-2.55 (m,4H),2.3 (t, 2H), 1.82-1.50 (m, 4H), 1.50-1.30 (m, 3H), 1.28-1.05 (m,8H), 0.98 (t, 3H), 0.75-0.60 (m, 3H).

EXAMPLE 7

N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-L-β-naphth-1-ylalanine ##STR12##

M.S., Calc'd: 569, Found: 569; NMR, δ=8.00-7.15 (m, 7H), 4.60-4.45 (m,2H), 3.71 (s, 2H), 3.65-3.50 (m, 2H), 3.40-2.98 (m, 4H), 2.70-2.42(m,4H), 2.21 (t, 2H), 1.70-1.45 (m, 2H), 1.40-0.96 (m, 7H), 0.92 (t,3H).

EXAMPLE 8

N- N-N-(4(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-L-β-naphth-2-ylalanine ##STR13##

M.S., Calc'd: 569, Found: 569; NMR, δ=7.65-7.03 (m, 7H), 4.60-4.45 (m,2H), 3.41 (s, 2H), 3.20-3.01 (m, 3H), 3.00-2.71 (m, 3H), 2.68-2.40(m,4H), (t, 2H), 1.68-1.42 (m, 3H), 1.25-0.85 (m, 6H), 0.71 (t, 3H).

EXAMPLE 9

N- N- N-(3-(piperidin-4-yl)propanoyol)-N-ethylglycyl!aspartyl! valine##STR14##

M.S., Calc'd: 547, Found: 457; NMR, δ=4.62 (m, 2H), 3.90 (s, 2H), 3.33(m, 4H), 2.66 (m, 4H), 2.37 (t, 2H), 2.16 (m, 1H), 2.03 (m, 1H), 1.78(m,2H), 1.44 (m, 2H), 1.20 (m, 2H), 1.00 (m, 3H), 0.78 (d, 6H).

EXAMPLE 10

N- N- N-(5-(piperidin-4-yl)pentanoyl)-N-ethylglycyl!aspartyl! valine##STR15##

M.S., Calc'd: 485, Found: 485; NMR, δ=4.20-4.05 (m, 2H), 3.92 (s, 2H),3.33 (m, 2H), 3.28-3.15 (m, 2H), 2.90-2.61 (m, 4H), 2.34 (t, 2H), 2.06(m, 1H), 1.85-1.70 (m, 2H), 1.55-1.32 (m, 3H), 1.30-1.12 (m, 6H), 1.06(t, 3H), 0.81 (d, 6H).

EXAMPLE 11

N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-L-β-cyclohexylalanine, ethyl ester ##STR16##A. β-cyclohexyl alanine (1.5 g) isdissolved in absolute ethanol (75 ml) and the solution cooled to 0° C.Thionyl chloride (1.1 ml) is added dropwise over a period of 10-15minutes, the solution allowed to warm to room temperature, and thenstirred at room temperature for about 18 hours. The reaction mixture isevaporated in vacuo, toluene is azeotroped twice from the residue, andthe residue is taken up into ethyl acetate. The ethyl acetate solutionis washed with water, 1N sodium hydroxide, water, brine, dried oversodium sulfate, filtered, and evaporated in vacuo to give β-cyclohexylalanine, ethyl ester.

B. Using essentially the procedures of Examples 1B through 1M(eliminating the aqueous sodium hydroxide hydrolysis of step 1L) givesthe desired product.

M.S., Calc'd: 553, Found: 553; NMR, δ=4.4 (1H, m); 4.1 (4H, q); 4.0 (2H,d; 3.2-3.5 (5H, m); 2.7-3.0 (5H, m); 2.4 (2H, t); 2.2 (1H, m); 1.9 (2H,d); 1.4-1.7 (7H, m); 0.7-1.4 (18H, m).

EXAMPLE 12

N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-L-β-cyclohexylalanine amide ##STR17##A. N-BOC-β-cyclohexylalanine (2.0 g.) andtriethyl amine (1.03 ml) aredissolved together in THF (100 ml) and thesolution cooled to -20° C. Isobutyl chloroformate (1.06 ml) is added andthe solution stirred at -20° C. for about 30 minutes. A saturatedsolution of ammonia in methanol (20 ml) is added and the solutionallowed to warm to room temperature and stirred at room temperature forabout 18 hours. The solution is evaporated in vacuo and the residuedissolved in ethyl acetate. The ethyl acetate solution is washed withwater, 5% HCl, saturated sodium bicarbonate solution, water, brine,dried over sodium sulfate, filtered and evaporated in vacuo to giveN-BOC-β-cyclohexylalanine amide.

B. N-BOC-β-cyclohexylalanine amide (2.0 g.) is dissolved in ethylacetate (100 ml) and HCl gas is bubbled through the solution and thesolution stirred at room temperature for about 18 hours. The solution isevaporated in vacuo and toluene azeotroped twice from the residue togive β-cyclohexylalanine amide as the hydrochloride.

C. Using essentially the procedures of Example 1B thru 1M (eliminatingthe aqueous sodium hydroxide hydrolysis of Example 1L) gives the desiredproduct.

M.S., Calc'd: 524, Found: 524; NMR, δ=8.4 (1H, d); 8.1 (1H, d); 4.2 (2H,q); 4.1 (1H, s); 3.9 (4H, q); 3.4 (2H, q); 3.3 (4H, d); 2.8-3.0 (6H, m);2.4 (2H, t); 2.2 (1H, m); 1.8 (4H, d); 1.4-1.7 (7H, m); 0.7-1.3 (10H,m).

EXAMPLE 13

N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-L-α-cyclohexylglycine ##STR18##A. A solution of α-phenylglycine, methyl ester,hydrochloride salt (1.0 g) in THF (25 ml) is cooled to 0° C. andtriethylamine (1.38 ml) is added. To this mixture is added a solution ofdi-tert-butyl dicarbonate (1.08 g) in THF (25 ml), and the mixtureallowed to warm to room temperature and stirred at room temperature forabout 18 hours. The solution is evaporated and the residue taken up intoethyl acetate (200 ml) and the organic solution washed with 1N HCl,saturated sodium bicarbonate solution, brine, dried over magnesiumsulfate, filtered, and evaporated in vacuo to giveN-BOC-α-phenylglycine, methyl ester.

B. N-BOC-α-phenylglycine, methyl ester (1.2 g) is dissolved in methanol(50 ml) containing acetic acid (1 ml). 5% rhodium on aluminum powder(0.60 g) is added and the mixture shaken under 50 psi hydrogen for about18 hours. The mixture is filtered, evaporated in vacuo, and the residuetaken up into ethyl acetate. The organic solution is washed with water,saturated sodium bicarbonate solution, water, brine, dried overmagnesium sulfate, filtered, evaporated in vacuo to giveN-BOC-α-cyclohexylglycine, methyl ester.

C. Using essentially the procedures of Examples 1B-1M, the desiredproduct is obtained.

M.S., Calc'd: 511, Found: 511; NMR, δ=4.62-4.55 (1H, m); 4.06 (2H, m);3.85 (2H, s); 3.30 (2H, q); 3.23-3.10 (2H, m); 2.85-2.55 (4H, m);2.30(2H, t); 1.83-1.60 (3H, m), 1.59-1.32 (8H, m); 1.30-0.75 (12H, m).

EXAMPLE 14

N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-L-β-decahydronaphth-1-ylalanine ##STR19##A. β-(1-naphthyl)alanine (2.0 g) is stirred in asaturated hydrogen chloride/methanol solution for about 2 hours at roomtemperature. The mixture is evaporated in vacuo and toluene azeotropedtwice from the residue. The residue is suspended in methylene chloride,N-methyl morpholine (1.02 ml) added, and the mixture cooled to 0° C.Di-tert-butyl dicarbonate (2.02 g) and 4-dimethylaminopyridine (DMAP)(0.8g) are added, the solution allowed to warm to room temperature, andstirredat room temperature for about 2 hours. The mixture is washed with5% HCl, water, dried over sodium sulfate, filtered, and evaporated invacuo to give N-BOC-β-(1 -naphthyl)alanine, methyl ester.

B. N-BOC-β-(1-naphthyl)alanine, methyl ester (2.0 g) and 5% rhodiumonalumina (1.0 g) are combined in methanol (50 ml) containing aceticacid (1.0 ml) and the mixture shaken under hydrogen at 50 psi for about18 hours. The mixture is filtered, evaporated in vacuo, and the residuetakenup into ethyl acetate. The organic solution is washed with water,5% sodiumbicarbonate solution, water, brine, dried over sodium sulfate,filtered, and evaporated in vacuo to give L-β-decahydronaphth-1-ylalanine, methyl ester.

C. Using essentially the procedures of Examples 1B through 1M, thedesired product is obtained.

M.S., Calc'd: 579, Found: 579; NMR, δ=4.1-4.3 (m, 1H), 3.8-4.1 (m, 2H),2.6-2.9 (m, 4H), 2.3 (m, 1H), 2.0 (m, 1H), 1.8 (d, 3H), 0.5-1.6 (m,33H),

EXAMPLE 15

2-cyclohexyl-N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-ethylamine##STR20##A. 2-phenylethylamine (2.0 g) is dissolved in methylenechloride and the solution cooled to 0° C. Di-tert-butyl dicarbonate (4.0g) and DMAP(0.4 g) are added. The solution is allowed to warm to roomtemperature and stirred at room temperature for about 18 hours. Thesolution is washed with 5% HCl, water, filtered and evaporated in vacuoto give N-BOC-2-phenylethylamine.

B. N-BOC-2-phenylethylamine (3.1 g) and 5% rhodium on alumina (1.1 g)are combined in methanol (40 ml) containing acetic acid (1.0 ml). Themixture is shaken under hydrogen at 50 psi for about 20 hours. Themixture is filtered, evaporated in vacuo, and the residue taken up intoethyl acetate. The organic solution is washed with water, 5% sodiumbicarbonate solution, water, brine, dried over sodium sulfate, filtered,and evaporated in vacuo to give N-BOC-2-cyclohexylethylamine.

C. Using essentially the procedures of Examples 1B through 1M(eliminating the aqueous sodium hydroxide hydrolysis of Example 1L), thedesired product is obtained.

M.S., Calc'd: 481, Found: 481; NMR, δ=3.9 (s, 2H), 3.35 (d, 4H), 3.25(d,4H), 2.6-2.9 (m, 8H), 2.35 (t, 2H), 2.15 (t, 1H), 1.8 (4H, d), 1,4-1.7(m, 7H), 0.9-1.3 (m, 12H), 0.7 (t, 2H).

EXAMPLE 16

N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-L-α-(2-cyclohexylethyl)glycine##STR21##A. Using essentially the procedures of Example 13A, above,N-BOC-α-(2-phenylethyl)glycine, methyl ester is prepared fromL-homophenylalanine.

B. Using essentially the procedure of Example 14B, above,N-BOC-α-(2-cyclohexylethyl)glycine, methyl ester is prepared fromN-BOC-α-(2-phenylethyl)glycine, methyl ester.

C. Using essentially the procedures of Examples 1B through 1M, thedesired product is obtained.

M.S., Calc'd: 539, Found: 539; NMR, δ=4.60-4.55 (m, 1H), 4.20-4.08 (m,1H), 3.85 (s, 2H), 3.29 (q, 2H), 3.25-3.12 (m, 2H), 2.84-2.55 (m,4H),2.29 (t, 2H), 1.83-1.65 (m, 2H), 1.63-1.32 (m, 10H), 1.28-0.81 (m,13H), 0.79-0.56 (m, 2H).

Using essentially the procedures of the above Examples, the followingcompounds are prepared from the appropriate starting materials.

EXAMPLE 17

N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-L-β-cis-decahydronaphth-2-ylalanine##STR22##

M.S., Calc'd: 579, Found: 579; NMR, δ=4.7 (m, 1H), 4.3 (m, 1H), 4.1 (d,2H), 3.3-3.7 (m, 5H), 2.6-2.8 (m, 5H), 2.5 (t, 2H), 2.3 (t, 1H), 1.9 (d,2H), 1.3-1.8 (m, 14H), 0.9-1.3 (m, 14H), 0.7-0.8 (m, 3H).

EXAMPLE 18

3-Adamant-1-ylpropyl-N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartate ##STR23##

M.S., Calc'd: 548, Found: 548; NMR (DMSO-d₆), δ=4.65-4.50 (m, 1H),4.05-3.85 (m, 4H), 3.35-3.15 (m, 4H), 2.90-2.50 (m, 4H), 2.30 (1, 2H),2.18 (t, 1H), 1.94 (d, 2H), 1.85-1.35 (m, 20H), 1.32-1.12 (m, 4H),1.10-0.90 (m, 5H).

EXAMPLE 19

N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-α-aminocyclohexanecarboxylicacid ##STR24##

M.S., Calc'd: 497, Found: 497; NMR, δ=4.60-4.55 (m, 1H), 4.05 (s, 1H),3.90 (s, 1H), 3.30 (q, 2H), 3.25-3.12 (m, 2H), 2.85-2.55 (m, 4H), 2.35(t, 1H), 2.11 (t, 1H), 1.90-1.70 (m, 4H), 1.68-1.55 (m, 2H), 1.53-1.32(m, 6H), 1.30-1.06 (m, 7H), 1.05-1.85 (m, 3H).

EXAMPLE 20

N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-β-cyclohexyl-D-alanine##STR25##

M.S., Calc'd: 525, Found: 525; NMR, δ=4.60-4.55 (m, 1H), 4.32-4.20 (m,1H), 4.05 (s, 1H), 3.85 (s, 1H), 3.32 (q, 2H), 3.25-3.12 (m, 2H),2.85-2.60 (m, 4H), 2.32 (t, 1H), 2.12 (t, 1H), 1.85-1.68 (m, 2H),1.60-1.32 (m, 10H), 1.28-0.60 (m, 13H).

EXAMPLE 21

N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-β-decahydronaphth-1-ylalanine##STR26##

M.S., Calc'd: 579, Found: 579; NMR, δ=4.1-4.3 (1H, m), 3.8-4.1 (2H, m),3.1-3.4 (4H, m), 2.6-2.9 (4H, m), 2.3 (1H, m), 2.0 (1H, m), 1.8 (3H, d),0.5-1.6 (33H, m).

EXAMPLE 22

N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-β-cyclohexylalanineethyl amide ##STR27##

M.S., Calc'd: 552, Found: 552; NMR, δ=4.55-4.45 (m, 1H), 4.20-4.06 (m,1H), 4.05-3.85 (m, 2H), 3.40-3.25 (m, 2H), 3.28-3.15 (m, 2H), 3.10-2.90(m, 2H), 2.88-2.55 (m, 4H), 2.40-2.25 (m, 1H), 2.20-2.05 (m, 1H),1.85-1.70 (m, 2H), 1.60-1.32 (m, 9H), 1.30-0.62 (m, 17H).

EXAMPLE 23

N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-β-cyclooctylalanine##STR28##

M.S., Calc'd: 553, Found: 553; NMR, δ=4.1-4.3 (1H, m), 3.8-4.1 (2H, m),3.1-3.4 (4H, m), 2.6-2.9 (4H, m), 2.3 (1H, m), 2.0 (1H, m), 1.8 (2H, d),0.5-1.6 (31H, m).

EXAMPLE 24

N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-α-cyclohexylmethylethanolamine##STR29##

M.S., Calc'd: 511, Found: 511; NMR, δ=4.60-4.45 (m, 1H), 4.10-3.75 (m,3H), 3.45-3.15 (m, 6H), 2.90-2.60 (m, 4H), 2.35 (t, 1H), 2.00-2.08(m,1H), 1.88-1.75 (m, 2H), 1.62-1.35 (m, 8H), 1.30-1.08 (m, 7H),1.10-0.60 (m,8H).

EXAMPLE 25

N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-β-cyclohexylmethylalanineamide ##STR30##

M.S., Calc'd: 538, Found: 538; NMR, δ=4.60-4.50 (m, 1H), 4.15-4.00 (m,1H), 4.00-3.80 (m, 2H), 3.35 (q, 2H), 3.30-3.15 (m, 2H), 2.90-2.62(m,4H), 2.35 (t, 1H), 2.15 (t, 1H), 1.88-1.75 (m, 2H), 1.65-1.40 (m,9H), 1.30-0.88 (m, 14H), 0.85-0.65 (m, 2H).

EXAMPLE 26

N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-β-adamant-1-ylalanine##STR31##

M.S., Calc'd: 577, Found: 577; NMR, δ=4.5-4.1 (1H, m), 4.1-4.2 (1H, m),3.8 (2H, d), 3.2 (2H, q), 3.1-3.1 (6H, m), 2.4-2.98 (5H, m), 2.3 (1H,m), 2.0 (1H, m), 1.8 (4H, m), 1.2-1.7 (16H, m), 1.0-1.2 (6H, m), 0.8-1.0(3H, m).

EXAMPLE 27

N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-β-(1,2,3,4)-tetrahydronaphth-5-ylalanine##STR32##

M.S., Calc'd: 573, Found: 573; NMR, δ=6.9 (d, 4H), 4.7 (m, 1H), 4.3 (m,1H), 4.1 (d, 2H), 3.3-3.7 (m, 6H), 2.6-3.1 (m, 12H), 2.5 (t, 2H), 2.3(t, 1H), 1.9 (d, 2H), 1.2-1.8 (m, 16H), 1.1 (t, 2H), 1.0 (t, 2H).

EXAMPLE 28

N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-β-(4-cyclohexyl)cyclohexylalanine##STR33##

M.S., Calc'd: 607, Found: 607; NMR, δ=4.2-4.3 (1H, m), 3.9-4.1 (2H, m),3.1-3.4 (5H, m), 2.6-2.9 (5H, m), 2.3 (1H, m), 2.0 (1H, m), 1.8 (3H, d),0.9-1.6 (32H, m), 0.7-0.8 (3H, m).

EXAMPLE 29

N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-β-cycloheptylalanine##STR34##

M.S., Calc'd: 539, Found: 539; NMR, δ=4.60-4.55 (m, 1H), 4.35-4.25 (m,1H), 4.08 (S, 1H), 3.92 (S, 1H), 3.35 (q, 2H), 3.33-3.20 (m, 2H),2.90-2.60 (m, 4H), 2.35 (t, 1H), 2.18 (t, 1H), 1.90-1.75 (m, 2H),1.70-1.10 (m, 22H), 1.10-0.85 (m, 3H).

EXAMPLE 30

N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-β-cyclooctylalanineamide ##STR35##

M.S., Calc'd: 551, Found: 551; NMR, δ=4.45 (dd, 1H, H-12), 4.18 (m, 1H,H-14), 3.89 (d, 1H, H-11), 3.69 (d, 1H, H-11), 3.31 (q, 2H, H-9),3.18(dt, 2H, H-1a), 2.74 (dt, 2H, H-le), 2.65 (dd, 2H, H-13), 2.25 (t,2H, H-8), 1.85-1.10 (m, 26H, H-3 through H-7 and H-15 through H-23),1.06 (t, 3H, H-10).

EXAMPLE 31

N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-α-cyclohexylpropylglycine##STR36##

M.S., Calc'd: 553, Found: 553; NMR, δ=4.70-4.60 (m, 1H), 4.30-4.15 (m,1H), 4.10 (S, 1H), 3.95 (S, 1H), 3.35 (q, 2H), 3.35-3.20 (m, 2H),2.90-2.60 (m, 4H), 2.40 (t, 1H), 2.15 (t, 1H), 1.90-1.75 (m, 2H),1.75-1.45 (m, 10H), 1.35-1.15 (m, 6H), 1.12-0.90 (m, 9H), 0.85-0.60 (m,2H).

EXAMPLE 32

N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-β-cyclooctylmethylalanine##STR37##

M.S., Calc'd: 567, Found: 567; NMR, δ=4.05-4.15 (m, 1H, 14), 3.75-4.00(m, 2H, 11 & 18), 3.10-3.30 (m, 4H, 19 & 26 eq), 2.50-2.80 (m, 4H, 15 &26 ax), 2.05-2.25 (m, 2H, 21), 0.75-1.75 (m, 31H, 1-10, 20 & 22-25).

EXAMPLE 33

N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-β-cyclopentylalanine##STR38##

M.S., Calc'd: 511, Found: 511; NMR, δ=4.7 (m, 1H), 4.3 (m, 1H), 4.1 (d,2H), 3.3-3.7 (m, 5H), 2.8 (t, 2H), 2.7 (m, 3H), 2.5 (t, 2H), 2.3 (t,1H), 1.9 (d, 2H), 1.0-1.8 (m, 16H).

EXAMPLE 34

N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-β-cyclohexylmethylalanineethyl ester ##STR39##

M.S., Calc'd: 567, Found: 567; NMR, δ=4.30-4.10 (m, 1H), 4.10-3.80 (m,3H), 3.35 (q, 2H), 3.30-3.15 (m, 2H), 2.90-2.60 (m, 4H), 2.40-2.10(t,2H), 1.90-1.70 (m, 2H), 1.65-1.40 (m, 10H), 1.35-0.85 (m, 18H),0.85-0.60 (m, 2H).

Compounds within the scope of the present invention inhibit plateletaggregation by inhibiting fibrinogen binding to activated platelets andother adhesive glycoproteins involved in platelet aggregation and bloodclotting and are useful in the prevention and treatment of thrombosisassociated with certain disease states, such as myocardial infarction,stroke, peripheral arterial disease and disseminated intravascularcoagulation in humans and other mammals.

The compounds of this invention can normally be administered orally orparenterally, in the treatment or prevention of thrombosis associateddisease states.

The compounds of this invention may be formulated for administration inanyconvenient way, and the invention includes within its scopepharmaceutical compositions containing at least one compound accordingto the invention adapted for use in human or veterinary medicine. Suchcompositions may be formulated in a conventional manner using one ormore pharmaceutically acceptable carriers or excipients. Suitablecarriers include diluents or fillers, sterile aqueous media and variousnon-toxic organic solvents. Thecompositions may be formulated in theform of tablets, capsules, lozenges, troches, hard candies, powders,aqueous suspensions, or solutions, injectable solutions, elixirs, syrupsand the like and may contain one or more agents selected from the groupincluding sweetening agents, flavoringagents, coloring agents andpreserving agents, in order to provide a pharmaceutically acceptablepreparation.

The particular carrier and the ratio of platelet aggregation andthrombus inhibiting compound to carrier are determined by the solubilityand chemical properties of the compounds, the particular mode ofadministration and standard pharmaceutical practice. For example,excipients such as lactose, sodium citrate, calcium carbonate anddicalcium phosphate and various disintegrants such as starch, alginicacidand certain complex silicates, together with lubricating agents suchas magnesium stearate, sodium lauryl sulphate and talc, can be used inproducing tablets. For a capsule form, lactose and high molecular weightpolyethylene glycols are among the preferred pharmaceutically acceptablecarriers. Where aqueous suspensions for oral use are formulated, thecarrier can be emulsifying or suspending agents. Diluents such asethanol,propylene glycol, glycerin and chloroform and their combinationscan be employed as well as other materials.

For parenteral administration, solutions or suspensions of thesecompounds in sesame or peanut oil or aqueous propylene glycol solutions,as well as sterile aqueous solutions of the soluble pharmaceuticallyacceptable saltsdescribed herein can be employed. Solutions of the saltsof these compoundsare especially suited for intramuscular andsubcutaneous injection purposes. The aqueous solutions, including thoseof the salts dissolved inpure distilled water, are also useful forintravenous injection purposes, provided that their pH is properlyadjusted, they are suitably buffered, they are made isotonic withsufficient saline or glucose and sterilized byheating ormicrofiltration.

The dosage regimen in carrying out the method of this invention is thatwhich insures maximum therapeutic response until improvement is obtainedand thereafter the minimum effective level which gives relief. Ingeneral,the oral dose may be between about 0.1 mg/kg and about 100mg/kg, preferably between about 0.1 mg/kg to 20 mg/kg, and mostpreferably between about 1 mg/kg and 20 mg/kg, and the i.v. dose about0.1 μg/kg to about 100 μg/kg, preferably between about 0.1 μg/kg to 50μg/kg, bearing in mind, of course, that in selecting the appropriatedosage in any specific case, consideration must be given to thepatient's weight, general health, age, and other factors which mayinfluence response to the drug. The drug may be administered orally 1 to4 times perday, preferably once to twice daily.

The following pharmacologic tests evaluate the inhibitory activity ofcompounds of the present invention on fibrinogen-mediated plateletaggregation, fibrinogen binding to thrombin-stimulated platelets, andinhibition of ADP-induced ex-vivo platelet aggregation, and results ofthese tests correlate to the in-vivo inhibitory properties of compoundsofthe present invention.

The Platelet Aggregation Assay is based on that described in Blood 66(4), 946-952 (1985). The Fibrinogen-Binding Assay is essentially that ofRuggeri, Z. M., et al., Proc. Natl. Acad. Sci. USA 83, 5708-5712 (1986)and Plow, E. F., et al., Proc. Natl. Acad. Sci., USA 82, 8057-8061(1985).The Inhibition of ADP-Induced ex-vivo Platelet Aggregation assayis based on that of Zucker, "Platelet Aggregation Measured by thePhotoelectric Method", Methods in Enzymology 169, 117-133 (1989).

Platelet Aggregation Assay Preparation of Fixed-Activated Platelets

Platelets are isolated from human platelet concentrates using thegel-filtration technique as described by Marguerie, G. A., et al., J.Biol. Chem. 254, 5357-5363 (1979) and Ruggeri, Z. M., et al., J. Clin.Invest. 72, 1-12 (1983). The platelets are suspended at a concentrationof2×10⁸ cells/ml in a modified calcium-free Tyrode's buffer containing127 mM sodium chloride, 2 mM magnesium chloride, 0.42 mM Na₂ HPO₄, 11.9mM NaHCO₃, 2.9 mM KCl, 5.5 mM glucose, 10 mMHEPES, at a pH of 7.35 and0.35% human serum albumin (HSA). These washed platelets are activated byaddition of human α-thrombin at a final concentration of 2 units/ml,followed by thrombin inhibitor I2581 at a final concentration of 40 μM.To the activated platelets is added paraformaldehyde to a finalconcentration of 0.50% and this incubated at room temperature for 30minutes. The fixed activated platelets are then collected bycentrifugation at 650×g for 15 minutes. The platelet pellets are washedfour times with the above Tyrode's-0.35% HSA buffer andresuspended to2×10⁸ cells/ml in the same buffer.

Platelet Aggregation Assay

The fixed activated platelets are incubated with a selected dose of thecompound to be tested for platelet aggregation inhibition for one minuteand aggregation initiated by addition of human fibrinogen to a finalconcentration of 250 μg/ml. A platelet aggregation profiler ModelPAP-4is used to record the platelet aggregation. The extent ofinhibition of aggregation is expressed as the percentage of the rate ofaggregation observed in the absence of inhibitor. IC₅₀, i.e., the amountof inhibitor required to reduce the aggregation rate by 50%, is thencalculated for each compound (see, for example, Plow, E. F., et al.,Proc.Natl. Acad. Sci., USA 82, 8057-8061 (1985)).

Fibrinogen-Binding Assay

Platelets are washed free of plasma constituents by the albumindensity-gradient technique of Walsh, P. N., et al., Br. J. Haematol.281-296 (1977), as modified by Trapani-Lombardo, V., et al., J. ClinInvest. 76, 1950-1958 (1985). In each experimental mixture platelets inmodified Tyrode's buffer (Ruggeri, Z. M., et al., J. Clin. Invest. 72,1-12 (1983)) are stimulated with human α-thrombin at 22°-25° C. for 10minutes (3.125×10¹¹ platelets per liter and thrombin at 0.1 NIHunits/ml). Hirudin is then added at a 25-fold excess (unit/unit) for 5minutes before addition of the ¹²⁵ I-labeled fibrinogen and the compoundto be tested. After these additions,the final platelet count in themixture is 1×10¹¹ /liter. After incubation for an additional 30 minutesat 22°-25° C., boundand free ligand are separated by centrifuging 50 μlof the mixture through 300 μl of 20% sucrose at 12,000×g for 4 minutes.The platelet pellet is then separated from the rest of the mixture todetermine platelet-bound radioactivity. Nonspecific binding is measuredinmixtures containing an excess of unlabeled ligand. When binding curvesare analyzed by Scatchard analysis, nonspecific binding is derived as afittedparameter from the binding isotherm by means of a computerizedprogram (Munson, P. J., Methods Enzymol. 92, 542-576 (1983)). Todetermine the concentration of each inhibitory compound necessary toinhibit 50% of fibrinogen binding to thrombin-stimulated platelets(IC₅₀), each compound is tested at 6 or more concentrations with ¹²⁵I-labeled fibrinogen held at 0.176 μmol/liter (60 μg/ml). The IC₅₀ isderived by plotting residual fibrinogen binding against the logarithm ofthe sample compound's concentration.

Inhibition of ADP-Induced ex-vivo Platelet Aggregation ExperimentalProtocol

Control blood samples are obtained 5-10 minutes prior to administrationof the test compound in mongrel dogs weighing from 10 to 20 kg. Thecompound is administered intragasticly, via aqueous gavage, or orally,via gelatin capsule. Blood samples (5 ml) are then obtained at 30 minuteintervals for3 hours, and at 6, 12, and 24 hours after dosing. Eachblood sample is obtained by venipuncture of the cephalic vein and iscollected directly into a plastic syringe containing one part 3.8%trisodium citrate to nine parts blood.

Ex vivo canine platelet aggregation

The blood samples are centrifuged at 1000 rpm for 10 minutes to obtainplatelet rich plasma (PRP). After removal of the PRP, the sample iscentrifuged for an additional 10 minutes at 2000 rpm to obtain plateletpoor plasma (PPP). Platelet count in the PRP is determined by using aCoulter Counter (Coulter Electronics, Hialeah, Fla.). If theconcentrationof platelets in the PRP is greater than 300,000platelets/μl, then the PRP is diluted with PPP to adjust the plateletcount to 300,000 platelets/μl. Aliquots of PRP (250 μl) are then placedin siliconized glass cuvettes (7.25×55 mm, Bio/Data Corp, Horsham, Pa.).Epinephrine (final concentration of 1 μM) is then added to the PRP,which is incubated for one minute at 37° C. A stimulator of plateletaggregation, ADP at a final concentration of 10 μM, is then added to thePRP. Platelet aggregation is monitored spectrophotometricallyutilizing alight transmission aggregometer (Bio/Data Platelet Aggregation Profiler,Model PAP-4, Bio/Data Corp, Horsham, Pa.). For compound to be tested,the rate of change (slope) of light transmittance and the maximum lighttransmittance (maximum aggregation) is recorded in duplicate. Plateletaggregation data are reported as the percent decrease (mean±SEM) inslope or maximum aggregation as compared to data obtainedfrom controlPRP, which is prepared from blood samples obtained prior toadministration of the test compound.

Compounds of the present invention exhibit marked activity in theforegoingtests and are considered useful in the prevention and treatmentof thrombosis associated with certain disease states. Antithromboticactivityin the ex vivo canine platelet aggregation assay is predictiveof such activity in humans (see, for example, Catalfamo, J. L., andDodds, W. Jean, "Isolation of Platelets from Laboratory Animals",Methods Enzymol. 169, Part A, 27 (1989)). Results of testing ofcompounds of the present invention by the above methods are presented inthe table below. Also presented in the table are comparative testresults for 4-4(piperidyl)butanoyl glycyl aspartyl tryptophan, i.e., thecompound disclosed in European Patent Application Publication No.0479,481.

    ______________________________________                      Inhibition of ADP-Induced            Inhibition                      ex-vivo Platelet Aggregation            of Fixed          % Inhibition of ex-vivo    Compound            Platelet          Platelet Aggregation    of Example            Aggregation                      Dose    After Oral Administration    Number  (IC.sub.50 μM)                      (mg/kg) 1 h  3 h  6 h  12 h 24 h    ______________________________________     2      0.027     10       90   60  <20     3      0.064     10       40  <20     9      0.77     1      0.026      5      100  100  100  88   42     4      0.72      10       90   50  <20     6      0.130     10       80   20     5      0.082     10      100   90   35    11      0.064      5      100  100   60  37    12      0.097      5      100  100  100  98   50     7      0.110      5       30  <20     8      0.068      5       30  <20    15      0.072      5       35   18    14      0.019      5      100  100  100  100  95    13                 5      100  100   25    17      0.096    18      0.104    19      3.41    20      1.37    21      0.055    22      0.09    23      0.078    24      0.129    25      0.046    26      0.029    27      0.119    28      0.048    29      0.032    30      0.07    31      0.053    32      0.052    33      0.047    34      0.074    (Compound            0.047     5        53  <20    of EPA    '481)    ______________________________________

One skilled in the art will readily appreciate that the presentinvention is well adapted to carry out the objects and obtain the endsand advantages mentioned, as well as those inherent therein. Thecompounds, compositions, and methods described herein are presented asrepresentativeof the preferred embodiments, or intended to be exemplaryand not intended as limitations on the scope of the present invention.Changes therein and other uses will occur to those of skill in the artwhich are encompassed within the spirit of the invention or defined bythe scope of the appendedclaims.

What is claimed is:
 1. A compound of the formula ##STR40## wherein: A is-H, amidino, or substituted amidino;B is alkyl, cycloalkyl,cycloalkylalkyl, alkylcycloalkyl, alkylcycloalkylalkyl, aryl, aralkyl,alkylaryl, or alkylaralkyl; ##STR41## where E is -H, F is -H, alkyl,hydroxymethyl, 1-hydroxyethyl, mercaptomethyl, 2-methylthioethyl,carboxymethyl, 2-carboxyethyl, aminocarbonylmethyl,2-aminocarbonylethyl, 4-aminobutyl, 3-aminopropyl, 3-guanidinopropyl,indol-3-ylmethyl, imidazol-3-ylmethyl, cycloalkyl, cycloalkylalkyl,cyclohexylcyclohexylmethyl, 1,2,3,4-tetrahydronaphth-5-ylmethyl,alkylcycloalkyl, alkylcycloalkylalkyl, aryl, substituted aryl, aralkyl,substitued aralkyl, heterocyclyl, substituted heterocyclyl,heterocyclylalkyl, substituted heterocyclylalkyl, wherein heterocyclylis pyridyl, pyrimidyl or pyrrolidyl, or, F taken together with E and thenitrogen and carbon atoms through which E and F are linked, forms a 4-,5-, 6-, or 7-membered azacycloalkane ring, wherein the azacycloalkanering is optionally substituted with hydroxy, G is alkyl, cycloalkyl,cycloalkylalkyl, alkylcycloalkyl, alkylcycloalkylalkyl, aryl,substituted aryl, aralkyl, substituted aralkyl, heterocyclyl,substituted heterocyclyl, heterocyclylalkyl, substitutedheterocyclylalkyl, wherein heterocyclyl is pyridyl, pyrimidyl orpyrrolidyl, OR¹, or NR¹ R², where R¹ and R² are independently -H, alkyl,cycloalkyl, cycloalkylalkyl, alkylcycloalkyl, alkylcycloalkylalkyl,aryl, aralkyl, alkylaryl, or alkylaralkyl, r is 0 or 1; R is H-, alkyl,aryl, or aralkyl; m is 1 to 5; n is 0 to 6; and p is 1 to 4; or apharmaceutically acceptable salt thereof.
 2. A compound of claim 1whereinF is -H, alkyl, hydroxymethyl, 1-hydroxyethyl, mercaptomethyl,2-methylthioethyl, carboxymethyl, 2-carboxyethyl, aminocarbonylmethyl,2-aminocarbonylethyl, 4-aminobutyl, 3-aminopropyl, 3-guanidinopropyl,phenylmethyl, 4-hydroxyphenylmethyl, indol-3-ylmethyl,imidazol-3-ylmethyl, cycloalkyl, cycloalkylalkyl,cyclohexylcyclohexylmethyl, 1,2,3,4-tetrahydronaphth-5-ylmethyl,alklylcycloalkyl, alkylcycloalkylalkyl, aryl, substituted aryl, aralkyl,substitued aralkyl, heterocyclyl, substituted heterocyclyl,heterocyclylalkyl, substituted heterocyclylalkyl, wherein heterocyclylis pyridyl, pyrimidyl or pyrrolidyl, or, F taken together with E and thenitrogen and carbon atoms through which E and F are linked, forms a 4-,5-, 6-, or 7-membered azacycloalkane ring, wherein the azacycloalkanering is optionally substituted with hydroxy, provided that when r is 1and G is -OR¹, then F is other than -H, methyl, isopropyl, isobutyl,sec-butyl, hydroxymethyl, 1-hydroxyethyl, mercaptomethyl,2-methylthioethyl, carboxymethyl, 2-carboxyethyl, aminocarbonylmethyl,2-aminocarbonylethyl, 4-aminobutyl, 3-aminopropyl, 3-guanidinopropyl,phenylmethyl, 4-hydroxyphenylmethyl, indol-3-ylmethyl, orimidazol-3-ylmethyl, or the group ##STR42##
 3. A compound of claim 2wherein F is -H, alkyl, hydroxymethyl, 1-hydroxyethyl, mercaptomethyl,2-methylthioethyl, carboxymethyl, 2-carboxyethyl, 4-aminobutyl,3-guanidinopropyl, cycloalkyl, cycloalkylalkyl, alkylcycloalkyl,alkylcycloalkylalkyl, aryl, substituted aryl, aralkyl, substitutedaralkyl, heterocyclyl, substituted heterocyclyl, heterocyclylalkyl,substituted heterocyclylalkyl, wherein heterocyclyl is pyridyl,pyrimidyl or pyrrolidyl, or, F taken together with E, and the nitrotgenand carbon atoms through which E and F are linked, forms a 4-, 5-, 6-,or 7-membered azacycloalkane ring, wherein the azacycloalkane ring isoptionally substituted with hydroxy, provided that when r is 1 and G is-OR¹, then F is other than -H, methyl, isopropyl, isobutyl, sec-butyl,hydroxymethyl, 1-hydroxyethyl, mercaptomethyl, 2-methylthioethylcarboxymethyl, 2-carboxyethyl, 4-aminobutyl, 3-guanidinopropyl,phenylmethyl, 4-hydroxyphenylmethyl, or the group ##STR43## or apharmaceutically acceptable salt thereof.
 4. A compound of claim 3wherein F is -H, alkyl, hydroxymethyl, 1-hydroxyethyl, mercaptomethyl,2-methylthioethyl, carboxymethyl, 2-carboxyethyl, 4-aminobutyl,3-guanidinopropyl, cycloalkyl, cycloalkylalkyl, alkylcycloalkyl,alkylcycloalkylalkyl, aryl, substituted aryl, aralkyl, substitutedaralkyl, or, F taken together with E, and the nitrogen and carbon atomsthrough which E and F are linked, forms a 4-, 5-, 6-, or 7-memberedazacycloalkane ring, wherein the azacycloalkane ring is optionallysubstituted with hydroxy, provided that when r is 1 and G is -OR¹, thenF is other than -H, methyl, isopropyl, isobutyl, sec-butyl,hydroxymethyl, 1-hydroxyethyl, mercaptomethyl, 2-methylthioethyl,carboxymethyl, 2-carboxyethyl, 4-aminobutyl, 3-guanidinopropyl,phenylmethyl, 4-hydroxyphenylmethyl, or the group ##STR44## or apharmaceutically acceptable salt thereof.
 5. A compound of claim 4wherein F is -H, alkyl, hydroxymethyl, 1-hydroxyethyl, mercaptomethyl,2-methylthioethyl, carboxy methyl, 2-carboxyethyl, 4-aminobutyl,3-guanidinopropyl, cycloalkyl, cycloalkylalkyyl, alkylcycloalkyl,alkylcycloalkylalkyl, or, F taken tog)ether with E, and the nitrogen andcarbon atoms through which E and F are linked, forms a 4-, 5-, 6-, or7-membered azacycloalkane ring, wherein the azacycloalkane ring, isoptionally substituted with hydroxy, provided that when r is 1 and G is-OR¹, then F is other than -H, methyl, isopropyl, isobutyl, sec-butyl,hydroxymethyl, 1-hydroxyethyl, mercaptomethyl, 2-methylthioethyl,carboxymethyl, 2-carboxyethyl, 4-aminobutyl, 3-guanidinopropyl, or thegroup ##STR45## or a pharmaceutically acceptable salt thereof.
 6. Acompound of claim 5 wherein B is alkyl, cycloalkyl, cycloalkylalkyl,alkylcycloalkyl, or alkylcycloalkylalkyl.
 7. A compound of claim 6wherein R¹ and R² are independently -H, alkyl, cycloalkyl,cycloalkylalkyl, alkylcycloalkyl or alkylcycloalkylalkyl.
 8. Apharmaceutical composition comprising an antithrombotic effective amountof a compound of claim 2 and a pharmaceutically acceptable carrier.
 9. Acompound of claim 1 of the formula ##STR46## wherein: A is -H oramidino,B is alkyl, cycloalkyl, cycloalkylalkyl, alkylcycloalkyl,alkylcycloalkylalkyl, aryl, aralkyl, alkylaryl, alkylaralkyl, J is -H,alkyl, cycloalkyl, cycloalkylalkyl, alkylcycloalkyl,alkylcycloalkylalkyl, aryl, substituted aryl, aralkyl, substituedaralkyl, cyclohexylcyclohexylmethyl, or1,2,3,4-tetrahydronaphth-5-ylmethyl, provided that when L is -OR¹, thenJ is other than -H, methyl, isopropyl, isobutyl, sec-butyl, phenylmethylor 4-hydroxyphenylmethyl, L is OR¹, or NR¹ R², where R¹ and R² areindependently -H, alkyl, cycloalkyl, cycloalkylalkyl, alkylcycloalkyl,alkylcycloalkylalkyl, aryl, aralkyl, alkylaryl, or alkylaralkyl, m is 1to 5, n is 2 to 6, and p is 1 or 2; or a pharmaceutically acceptablesalt thereof.
 10. A compound of claim 9 whereinA is -H, B is alkyl,cycloalkyl, cycloalkylalkyl, alkylcycloalkyl, or alkylcycloalkylalkyl, Jis -H, alkyl, cycloalkyl, cycloalkylalkyl, alkylcycloalkyl,alkylcycloalkylalkyl, or cyclohexylcyclohexylmethyl, provided that whenL is -OR¹, then J is other than -H, methyl, isopropyl, isobutyl,sec-butyl, phenylmethiyl or 4-hydroxyplhenylmethyl, m is 3, and n is 3or 4, or a pharmaceutically acceptable salt thereof.
 11. A compound ofclaim 10 whereinA is -H, B is alkyl, J is alkyl, cycloalkyl, orcycloalkylalkyl, or cyclohexylcyclohexylmethyl, provided that when L is-OR¹, then J is other than methyl, isopropyl, isobutyl, or sec-butyl, R¹and R² are independently -H, alkyl, cycloalkyl, cycloalkylalkyl,alkylcycloalkyl, or alkylcycloalkylalkyl, m is 3, n is 3 or 4, and p is1, or a pharmaceutically acceptable salt thereof.
 12. A pharmaceuticalcomposition comprising an antithrombotic effective amount of a compoundof claim 9 and a pharmaceutically acceptable carrier.
 13. Apharmaceutical composition comprising an antithrombotic effective amountof a compound of claim 1 and a pharmaceutically acceptable carrier. 14.A compound of claim 1 of the formula ##STR47## wherein: A is -H,amidino, or substituted amidino,B is alkyl, cycloalkyl, cycloalkylalkyl,alkylcycloalkyl, alkylcycloalkylalkyl, aryl, aralkyl, alkylaryl, oralkylaralkyl; ##STR48## where E is -H, F is -H, methyl, isopropyl,isobutyl, sec-butyl, hydroxymethyl, 1-hydroxyethyl, mercaptomethyl,2-methylthioethyl, carboxymethyl, 2-carboxyethyl, aminocarbonylmethyl,2-aminocarbonylethyl, 4-aminobutyl, 3-aminopropyl, 3-guanidinopropyl,phenylmethyl, 4-hydroxyphenylmethyl, indol-3-ylmethyl,imidazol-3-ylmethyl, or the group ##STR49## G is -OR¹ wherein R¹ is -H,alkyl, cycloalkyl, cycloalkylalkyl, alkylcycloalkyl, oralkylcycloalkylalkyl, r is 1; R is -H, alkyl, aryl, or aralkyl; m is 1to 5; n is 0 to 6; and p is 1 to 4; or a pharmaceutically acceptablesalt thereof.
 15. A compound of claim 14 wherein F is -H, methyl,isopropyl, isobutyl, sec-butyl, hydroxymethyl, 1-hydroxyethyl,mercaptomethyl, 2-methylthioethyl, carboxymethyl, 2-carboxyethyl,4-aminobutyl, 3-guanidinopropyl, or the group ##STR50## or apharmaceutically acceptable salt thereof.
 16. A compound of claim 15wherein B is alkyl, cycloalkyl, cycloalkylalkyl, alkylcycloalkyl, oralkylcycloalkylalkyl.
 17. A pharmaceutical composition comprising anantithrombotic effective amount of a compound of claim 14 and apharmaceutically acceptable carrier.
 18. A compound of claim 1 of theformula ##STR51## wherein: A is -H or amidino:B is alkyl, cycloalkyl,cycloalkylalkyl, alkylcycloalkyl, alkylcycloalkylalkyl, aryl, aralkyl,alkylaryl, or alkylaralkyl; J is -H, methyl, isopropyl, isobutyl,sec-butyl; L is -OR¹ wherein R¹ is -H, alkyl, cyeloalkyl,cycloalkylalkyl, alkylcycloalkyl, or alkylcycloalkylalkyl, m is 1 to 5;n is 2 to 6; and p is 1 or 2; or a pharmaceutically acceptable saltthereof.
 19. A compound of claim 18 whereinA is -H; B is alkyl,cycloalkyl, cycloalkylalkyl, alkylcycloalkyl, alkylcycloalkylalkyl; m is3; and n is 3 or 4; or a pharmaceutically acceptable salt thereof.
 20. Acompound of claim 19 whereinB is alkyl; L is -OR¹ wherein R¹ is -H,alkyl, cycloalkyl, cycloalkylalkyl or alkylcycloalkylalkyl; m is 3; n is3 or 4; and p is 1; or a pharmaceutically acceptable salt thereof.
 21. Apharmaceutical composition comprising an antithrombotic effective amountof a compound of claim 18 and a pharmaceutically acceptable carrier. 22.A compound which isN- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl! valine, N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-D-valine, N- N-N-(3-(piperidin-4-yl)propanoyl)-N-ethylglycyl!aspartyl! valine, N- N-N-(5-(piperidin-4-yl)pentanoyl)-N-ethylglycyl!aspartyl! valine, N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-L-α-cyclohexylglycine, N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-β-cyclohexylalanine,N- N- N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl! norleucine,N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-L-α-(2,2-dimethyl)prop3-ylglycine, N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-L-β-cis-decahydronaphth-2-ylalanine,N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-α-aminocyclohexanecarboxylicacid, N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-β-cyclohexyl-D-alaniine,N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-β-decahydronaphth-1-ylalanine,N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-β-cyclooctylalamine,N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspattyl!-β-adamant-1-ylalanine,N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-β-(4-cyclohexyl)cyclohexylalanine,N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-β-cycloheptylalanine,N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-α-cyclohexylpropylglycine,N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-β-cyclooctylmethylalanine,N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-β-cyclopentylalanineN- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-L-β-decahydronaphth-1-ylalanine, N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-L-α-(2-cyclohexylethyl)glycine,N- N- N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!phenylalanine, N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-β-(1,2,3,4)-tetrahydronaphth-5-ylalanine,N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-L-β-naphth-1-ylalanine, N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-L-β-naphth-2-ylalanine, N- N-N-(4-(pipenidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-L-β-cyclohexylalanine amide, N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-β-cyclooctylalanineamide, N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-β-cyclohexylmethylalanineamide, N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-β-cyclohexylalanineethyl amiide, N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-L-β-cyclohexylalanine, ethyl ester, N- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-β-cyclohexylmethylalanineethyl ester, 3-Adamant-1-ylpropyl-N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartate, 2-cyclohexyl-N-N- N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-ethylamine, orN- N-N-(4-(piperidin-4-yl)butanoyl)-N-ethylglycyl!aspartyl!-α-cyclohexylmethylethanolamine,or a pharmaceutically acceptable salt thereof.
 23. A pharmaceuticalcomposition comprising an antithrombotic effective amount of a compoundof claim 22 and a phanmaceutically acceptable carrier.
 24. A method forthe prevention or treatment of thrombosis in a mammal in need of suchtherapy comprising the administration of a therapeutically effectiveamount of a compound of claim
 22. 25. A method for the prevention ortreatment of thrombosis in a mammal in need of such therapy comprisingthe administration of a therapeutically effective amount of thecomposition of claim
 23. 26. A method for the prevention or treatment ofthrombosis in a mammal in need of such therapy comprising theadministration of a therapeutically effective amount of a compound ofclaim
 2. 27. A method for the prevention or treatment of thrombosis in amammal in need of such therapy comprising the administration of atherapeutically effective amount of the composition of claim
 8. 28. Amethod for the treatment of thrombosis in a mammal in need of suchtherapy comprising the administration of a therapeutically effectiveamount of a compound of claim
 9. 29. A method for the prevention ortreatment of thrombosis in a mammal in need of such therapy comprisingthe administration of a therapeutically effective amount of thecomposition of claim
 14. 30. A method for the prevention or treatment ofthrombosis in a mammal in need of such therapy comprising theadministration of a therapeutically effective amount of the compositionof claim
 13. 31. A method for the treatment of thrombosis in a mammal inneed of such therapy comprising the administration of a therapeuticallyeffective amount of a compound of claim
 1. 32. A method for theprevention or treatment of thrombosis in a mammal in need of suchtherapy comprising the administration of a therapeutically effectiveamount of a compound of claim
 14. 33. A method for the prevention ortreatment of thrombosis in a mammal in need of such therapy comprisingthe administration of a therapeutically effective amount of thecomposition of claim
 17. 34. A method for the prevention or treatment ofthrombosis in a mammal in need of such therapy comprising theadministration of a therapeutically effective amount of a compound ofclaim
 18. 35. A method for the prevention or treatment of thrombosis ina mammal in need of such therapy comprising the administration of atherapeutically effective amount of the composition of claim 21.